1000 resultados para nifB point-mutated


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Finding an appropriate linking method to connect different dimensional element types in a single finite element model is a key issue in the multi-scale modeling. This paper presents a mixed dimensional coupling method using multi-point constraint equations derived by equating the work done on either side of interface connecting beam elements and shell elements for constructing a finite element multiscale model. A typical steel truss frame structure is selected as case example and the reduced scale specimen of this truss section is then studied in the laboratory to measure its dynamic and static behavior in global truss and local welded details while the different analytical models are developed for numerical simulation. Comparison of dynamic and static response of the calculated results among different numerical models as well as the good agreement with those from experimental results indicates that the proposed multi-scale model is efficient and accurate.

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The conversion of tamarind seeds into pyrolytic oil by fixed bed fire-tube heating reactor has been taken into consideration in this study. The major components of the system were fixed bed fire-tube heating reactor, liquid condenser and collectors. The raw and crushed tamarind seed in particle form was pyrolized in an electrically heated 10 cm diameter and 27 cm high fixed bed reactor. The products are oil, char and gases. The parameters varied were reactor bed temperature, running time, gas flow rate and feed particle size. The parameters were found to influence the product yields significantly. The maximum liquid yield was 45 wt% at 4000C for a feed size of 1.07cm3 at a gas flow rate of 6 liter/min with a running time of 30 minute. The pyrolysis oil was obtained at these optimum process conditions were analyzed for physical and chemical properties to be used as an alternative fuel.

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This report reviews the use of point-to-point speed enforcement internationally in order to provide principles for better practice for its use in Australia and New Zealand. Point-to-point enforcement is a relatively new technological approach to speed enforcement which involves measuring the average speed of vehicles passing a series of cameras, by using automatic number plate recognition (ANPR) and other technologies. The approach has been implemented or trialled in a number of countries including Australia, New Zealand, the United Kingdom, the Netherlands, Austria, Italy, Switzerland and France. The major research activities were undertaken for the preparation of this report included: (1) an extensive review of the international literature; (2) stakeholder consultation with international and domestic organisations. To date, there have been no formal evaluations of point-to- point speed enforcement in Australia or New Zealand.

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Protease-activated receptor-2 (PAR2) is a G protein coupled receptor (GPCR) that is activated by proteolytic cleavage of its amino terminal domain by trypsin-like serine proteases. Cleavage of this receptor exposes a neoepitope, termed the tethered ligand (TL), which binds intramolecularly within the receptor to stimulate signal transduction via coupled G proteins. PAR2-mediated signal transduction is also experimentally stimulated by hexapeptides (agonist peptides; APs) that are homologous to the TL sequence. Due to the irreversible nature of PAR2 proteolysis, downstream signal transduction is tightly regulated. Following activation, PAR2 is rapidly uncoupled from downstream signalling by the post-translational modifications phosphorylation and ubiquination which facilitate interactions with â- arrestin. This scaffolding protein couples PAR2 to the internalisation machinery initiating its desensitisation and trafficking through the early and late endosomes followed by receptor degradation. PAR2 is widely expressed in mammalian tissues with key roles for this receptor in cardiovascular, respiratory, nervous and musculoskeletal systems. This receptor has also been linked to pathological states with aberrant expression and signalling noted in several cancers. In prostate cancer, PAR2 signalling induces migration and proliferation of tumour derived cell lines, while elevated receptor expression has been noted in malignant tissues. Importantly, a role for this receptor has also been suggested in prostate cancer bone metastasis as coexpression of PAR2 and a proteolytic activator has been demonstrated by immunohistochemical analysis. Based on these data, the primary focus of this project has been on two aspects of PAR2 biology. The first is characterisation of cellular mechanisms that regulate PAR2 signalling and trafficking. The second aspect is the role of this receptor in prostate cancer bone metastasis. In addition, to permit these studies, it was first necessary to evaluate the specificity of the commercially available anti-PAR2 antibodies SAM11, C17, N19 and H99. The evaluation of the four commercially available antibodies was assessed using four techniques: immunoprecipitation; Western blot analysis; immunofluorescence; and flow cytometry. These approaches demonstrated that three of the antibodies efficiently detect ectopically expressed PAR2 by each of these techniques. A significant finding from this study was that N19 was the only antibody able to specifically detect N-glycosylated endogenous PAR2 by Western blot analysis. This analysis was performed on lysates from prostate cancer derived cell lines and tissue derived from wildtype and PAR2 knockout mice. Importantly, further evaluation demonstrated that this antibody also efficiently detects endogenous PAR2 at the cell surface by flow cytometry. The anti-PAR2 antibody N19 was used to explore the in vitro role of palmitoylation, the post-translational addition of palmitate, in PAR2 signalling, trafficking, cell surface expression and desensitization. Significantly, use of the palmitoylation inhibitor 2-bromopalmitate indicated that palmitate addition is important in trafficking of PAR2 endogenously expressed by prostate cancer cell lines. This was supported by palmitate labelling experiments using two approaches which showed that PAR2 stably expressed by CHO cells is palmitoylated and that palmitoylation occurs on cysteine 361. Another key finding from this study is that palmitoylation is required for optimal PAR2 signalling as Ca2+ flux assays indicated that in response to trypsin agonism, palmitoylation deficient PAR2 is ~9 fold less potent than wildtype receptor with a reduction of about 33% in the maximum signal induced via the mutant receptor. Confocal microscopy, flow cytometry and cell surface biotinylation analyses demonstrated that palmitoylation is required for efficient cell surface expression of PAR2. Importantly, this study also identified that palmitoylation of this receptor within the Golgi apparatus is required for efficient agonist-induced rab11amediated trafficking of PAR2 to the cell surface. Interestingly, palmitoylation is also required for receptor desensitization, as agonist-induced â-arrestin recruitment and receptor degradation were markedly reduced in CHO-PAR2-C361A cells compared with CHO-PAR2 cells. Collectively, these data provide new insights on the life cycle of PAR2 and demonstrate that palmitoylation is critical for efficient signalling, trafficking, cell surface localization and degradation of this receptor. This project also evaluated PAR2 residues involved in ligand docking. Although the extracellular loop (ECL)2 of PAR2 is known to be required for agonist-induced signal transduction, the binding pocket for receptor agonists remains to be determined. In silico homology modelling, based on a crystal structure for the prototypical GPCR rhodopsin, and ligand docking were performed to identify PAR2 transmembrane (TM) amino acids potentially involved in agonist binding. These methods identified 12 candidate residues that were mutated to examine the binding site of the PAR2 TL, revealed by trypsin cleavage, as well as of the soluble ligands 2f-LIGRLO-NH2 and GB110, which are both structurally based on the AP SLIGRLNH2. Ligand binding was evaluated from the impact of the mutated residues on PAR2-mediated calcium mobilisation. An important finding from these experiments was that mutation of residues Y156 and Y326 significantly reduced 2f-LIGRLO-NH2 and GB110 agonist activity. L307 was also important for GB110 activity. Intriguingly, mutation of PAR2 residues did not alter trypsin-induced signalling to the same extent as for the soluble agonists. The reason for this difference remains to be further examined by in silico and in vitro experimentation and, potentially, crystal structure studies. However, these findings identified the importance of TM domains in PAR2 ligand docking and will enhance the design of both PAR2 agonists and potentially agents to inhibit signalling (antagonists). The potential importance of PAR2 in prostate cancer bone metastasis was examined using a mouse model. In patients, prostate cancer bone metastases cause bone growth by disrupting bone homeostasis. In an attempt to mimic prostate cancer growth in bone, PAR2 responsive 22Rv1 prostate cancer cells, which form mixed osteoblastic and osteolytic lesions, were injected into the proximal aspect of mouse tibiae. A role for PAR2 was assessed by treating these mice with the recently developed PAR2 antagonist GB88. As controls, animals bearing intra-tibial tumours were also treated with vehicle (olive oil) or the prostate cancer chemotherapeutic docetaxel. The effect of these treatments on bone was examined radiographically and by micro-CT. Consistent with previous studies, 22Rv1 tumours caused osteoblastic periosteal spicule formation and concurrent osteolytic bone loss. Significantly, blockade of PAR2 signalling reduced the osteoblastic and osteolytic phenotype of 22Rv1 tumours in bone. No bone defects were detected in mice treated with docetaxel. These qualitative data will be followed in the future by quantitative micro-CT analysis as well as histology and histomorphometry analysis of already collected tissues. Nonetheless, these preliminary experiments highlight a potential role for PAR2 in prostate cancer growth in bone. In summary, in vitro studies have defined mechanisms regulating PAR2 activation, downstream signalling and trafficking and in vivo studies point to a potential role for this receptor in prostate cancer bone metastasis. The outcomes of this project are that a greater understanding of the biology of PAR2 may lead to the development of strategies to modulate the function of this receptor in disease.

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This paper presents a reactive collision avoidance method for small unmanned rotorcraft using spherical image-based visual servoing. Only a single point feature is used to guide the aircraft in a safe spiral like trajectory around the target, whilst a spherical camera model ensures the target always remains visible. A decision strategy to stop the avoidance control is derived based on the properties of spiral like motion, and the effect of accurate range measurements on the control scheme is discussed. We show that using a poor range estimate does not significantly degrade the collision avoidance performance, thus relaxing the need for accurate range measurements. We present simulated and experimental results using a small quad rotor to validate the approach.

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Recombinant human papillomavirus (HPV) virus-like particles (VLPs) made from the major capsid protein L1 are promising vaccine candidates for use as vaccines against genital and other HPV infections, and particularly against HPV-16. However, HPV-16 genotype variants have different binding affinities for neutralising mouse Mabs raised against HPV-16 L1 VLPs. This paper analyses, using a panel of well-characterised Mabs, the effects on the antigenicity of various C- and N-terminal deletants of HPV-16 L1 made in insect cells via recombinant baculovirus, of an A → T mutation at residue 266 (A266T), and of a C → G mutation at conserved position 428 (C428G). The effects of these changes on assembly of the variant L1s were studied by electron microscopy. Binding of Mab H16:E70 to A266T was reduced by almost half in comparison to wild type L1. Retention of the C-terminal region 428-483 was critical for the binding of conformation-specific Mabs (H16:V5, H16:E70, H16:U4 and H16:9A) whereas deletion of the nuclear localisation signal (NLS) or the C428G mutation or an N-terminal deletion (residues 2-9) did not affect the antigenicity. The N-terminal deletion resulted in a mixed population of 30 and 55 nm VLPs, which differs from the same construct expressed in Escherichia coli, whereas pentamer aggregates resulted from deletion of the 428-465 region or the C428G mutation. The results have implications both for considering use of single-genotype HPV vaccines, and for design of novel second-generation vaccines. © 2006 Elsevier B.V. All rights reserved.

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This paper is concerned with recent advances in the development of near wall-normal-free Reynolds-stress models, whose single point closure formulation, based on the inhomogeneity direction concept, is completely independent of the distance from the wall, and of the normal to the wall direction. In the present approach the direction of the inhomogeneity unit vector is decoupled from the coefficient functions of the inhomogeneous terms. A study of the relative influence of the particular closures used for the rapid redistribution terms and for the turbulent diffusion is undertaken, through comparison with measurements, and with a baseline Reynolds-stress model (RSM) using geometric wall normals. It is shown that wall-normal-free rsms can be reformulated as a projection on a tensorial basis that includes the inhomogeneity direction unit vector, suggesting that the theory of the redistribution tensor closure should be revised by taking into account inhomogeneity effects in the tensorial integrity basis used for its representation.

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This paper illustrates the complexity of pointing as it is employed in a design workshop. Using the method of interaction analysis, we argue that pointing is not merely employed to index, locate, or fix reference to an object. It also constitutes a practice for reestablishing intersubjectivity and solving interactional trouble such as misunderstandings or disagreements by virtue of enlisting something as part of the participants’ shared experience. We use this analysis to discuss implications for how such practices might be supported with computer mediation, arguing for a “bricolage” approach to systems development that emphasizes the provision of resources for users to collaboratively negotiate the accomplishment of intersubjectivity ra- ther than systems that try to support pointing as a specific gestural action.

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Five basalt samples from the Point Sal ophiolite, California, were examined using HRTEM and AEM in order to compare observations with interpretations of XRD patterns and microprobe analyses. XRD data from ethylene-glycol-saturated samples indicate the following percentages of chlorite in mixed-layer chlorite-smectite identified for each specimen: (i) L2036 almost-equal-to 50%, (ii) L2035 almost-equal-to 70 and 20%, (iii) 1A-13 almost-equal-to 70%, (iv) 1B-42 almost-equal-to 70%, and (v) 1B-55 = 100%. Detailed electron microprobe analyses show that 'chlorite' analyses with high Si, K, Na and Ca contents are the result of interlayering with smectite-like layers. The Fe/(Fe + Mg) ratios of mixed-layer phyllosilicates from Point Sal samples are influenced by the bulk rock composition, not by the percentage of chlorite nor the structure of the phyllosilicate. Measurements of lattice-fringe images indicate that both smectite and chlorite layers are present in the Point Sal samples in abundances similar to those predicted with XRD techniques and that regular alternation of chlorite and smectite occurs at the unit-cell scale. Both 10- and 14-angstrom layers were recorded with HRTEM and interpreted to be smectite and chlorite, respectively. Regular alternation of chlorite and smectite (24-angstrom periodicity) occurs in upper lava samples L2036 and 1A-13, and lower lava sample 1B-42 for as many as seven alternations per crystallite with local layer mistakes. Sample L2035 shows disordered alternation of chlorite and smectite, with juxtaposition of smectite-like layers, suggesting that randomly interlayered chlorite (< 0.5)-smectite exists. Images of lower lava sample 1B-55 show predominantly 14-angstrom layers. Units of 24 angstrom tend to cluster in what may otherwise appear to be disordered mixtures, suggesting the existence of a corrensite end-member having thermodynamic significance.

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The International Baccalaureate Diploma is an independent, globally available curriculum currently enjoying rapid uptake in government systems as an alternative curriculum. This paper explores the logic of its consumption in three case study schools across different states of Australia, and the relational ‘points of difference’ it creates in each local context and its curricular market. The analysis uses a typology of goods to describe the nature and dynamics of the IBD’s glocalised ecology of in each site. The conclusion argues the success of the IBD as a curricular alternative risks eroding its appeal as a positional good.

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Video-based training combined with flotation tank recovery may provide an additional stimulus for improving shooting in basketball. A pre-post controlled trial was conducted to assess the effectiveness of a 3 wk intervention combining video-based training and flotation tank recovery on three-point shooting performance in elite female basketball players. Players were assigned to an experimental (n=10) and control group (n=9). A 3 wk intervention consisted of 2 x 30 min float sessions a week which included 10 min of video-based training footage, followed by a 3 wk retention phase. A total of 100 three-point shots were taken from 5 designated positions on the court at each week to assess three-point shooting performance. There was no clear difference in the mean change in the number of successful three-point shots between the groups (-3%; ±18%, mean; ±90% confidence limits). Video-based training combined with flotation recovery had little effect on three-point shooting performance.

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Objective: To determine the impact of a free-choice diet on nutritional intake and body condition of feral horses. Animals: Cadavers of 41 feral horses from 5 Australian locations. Procedures: Body condition score (BCS) was determined (scale of 1 to 9), and the stomach was removed from horses during postmortem examination. Stomach contents were analyzed for nutritional variables and macroelement and microelement concentrations. Data were compared among the locations and also compared with recommended daily intakes for horses. Results: Mean BCS varied by location; all horses were judged to be moderately thin. The BCS for males was 1 to 3 points higher than that of females. Amount of protein in the stomach contents varied from 4.3% to 14.9% and was significantly associated with BCS. Amounts of water-soluble carbohydrate and ethanol-soluble carbohydrate in stomach contents of feral horses from all 5 locations were higher than those expected for horses eating high-quality forage. Some macroelement and microelement concentrations were grossly excessive, whereas others were grossly deficient. There was no evidence of ill health among the horses. Conclusions and Clinical Relevance: Results suggested that the diet for several populations of feral horses in Australia appeared less than optimal. However, neither low BCS nor trace mineral deficiency appeared to affect survival of the horses. Additional studies on food sources in these regions, including analysis of water-soluble carbohydrate, ethanol-soluble carbohydrate, and mineral concentrations, are warranted to determine the provenance of such rich sources of nutrients. Determination of the optimal diet for horses may need revision.

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This work experimentally examines the performance benefits of a regional CORS network to the GPS orbit and clock solutions for supporting real-time Precise Point Positioning (PPP). The regionally enhanced GPS precise orbit solutions are derived from a global evenly distributed CORS network added with a densely distributed network in Australia and New Zealand. A series of computational schemes for different network configurations are adopted in the GAMIT-GLOBK and PANDA data processing. The precise GPS orbit results show that the regionally enhanced solutions achieve the overall orbit improvements with respect to the solutions derived from the global network only. Additionally, the orbital differences over GPS satellite arcs that are visible by any of the five Australia-wide CORS stations show a higher percentage of overall improvements compared to the satellite arcs that are not visible from these stations. The regional GPS clock and Uncalibrated Phase Delay (UPD) products are derived using the PANDA real time processing module from Australian CORS networks of 35 and 79 stations respectively. Analysis of PANDA kinematic PPP and kinematic PPP-AR solutions show certain overall improvements in the positioning performance from a denser network configuration after solution convergence. However, the clock and UPD enhancement on kinematic PPP solutions is marginal. It is suggested that other factors, such as effects of ionosphere, incorrectly fixed ambiguities, may be the more dominating, deserving further research attentions.

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This thesis consists of the novel Brolga and an exegesis examining in what ways the ideas of katabasis and deterritorialisation inform an understanding of descent narratives in contemporary Australian outback fiction. When writing the creative piece, it was observed that Joseph Campbell’s Hero’s Journey was an imprecise model for my manuscript and indeed for many of the contemporary novels I had read written in similar outback settings. On analysis a better fit lies in the idea of a heroic journey from which there is no clear return from the underworld. This narrative form is defined in this thesis as a katabatic narrative. To unpack this narrative trope, the inverse of territoriality, deterritorialisation, is used as a lens to examine the complex thematic and symbolic resonances of the outback in both Brolga and analogous works of contemporary outback fiction.